Part B | 10.11
302 Part B Tools and Methods in Marine Biotechnology
other things, the improvement of the growth rate or
the increase in thermotolerance. Growth enhancement
strategies have been successfully applied to several
fish species, such as carp, catfish, tilapia, and salmon,
through the incorporation of homologous or heterologous growth hormone (GH) genes or cDNAs. The
monitoring of gene expression in genetically modified
marine organisms is thus a crucial step in the optimization of the use of such organisms in aquaculture
and seafood technology. It should also be noted that
the cross-effect of transgene expression on the biosynthesis of host proteins in genetically improved fish
species is another relevant issue in which proteomics
will surely play a significant role soon. In both issues,
proteomics represents a valuable tool of remarkable applied interest.
10.10 Present Status and Future Trends of Proteomics
in Marine Biotechnology
Proteomic tools, mainly 2-D PAGE (polyacrylamide
gel electrophoresis) and mass spectrometry, represent
a robust analytical strategy for the phenotypical investigation of marine organisms of industrial interest. These
tools afford specific information about expression patterns, post-translational modifications, or protein interaction mechanisms. This information is extremely
useful for the optimization of specific biotechnological processes. It should be stressed that high-throughput
proteomics techniques have recently been coupled with
newer bioinformatic tools, thus affording more robust
and powerful strategies for both protein identification and characterization purposes. Recently, proteomic
tools were successfully applied for the isolation and
characterization of a calponin-like protein from the
mussel Mytilus galloprovincialis 68 kDa polypeptide,
a polypeptide involved in muscle contraction processes.
The protein, isolated from the retractor muscle of this
mollusc, was successfully separated from actin by 2-D
PAGE and was identified by N-terminal amino acid
sequence analysis. Likewise, the investigation of mussel adhesive proteins which have been reported to
harbor an extraordinary biotechnological potential as
a biodegradable adhesive material is another promising field in which proteomics may play a significant
role. It is noteworthy that the protein can be readily obtained from mussels, a marine source characterized by
their wide availability and low price. Thus, proteomic
tools should play a significant role when undertaking
optimization strategies aimed at achieving the highest yields of such proteins, mainly by elucidating the
biochemical modifications to be considered in the process design step. The investigation of marine bacteria
by proteomic techniques is also still in its early days.
Although the protein profiles of the thermophilic microorganism Pyrococcus abyssi were investigated when
the organism was subjected to different levels of pressure and temperature, other authors applied 2-D PAGE
to investigate the level of protein biosynthesis and expression in Sphingomonas sp. cultivated under different
conditions. Earlier reports considered high-resolution
2-D PAGE to elucidate the mechanisms of UV protection in cyanobacteria, concluding that this technique
had greater power of resolution than conventional SDSPAGE (SDS: sodium dodecyl sulfate) to detect proteome changes. The relationship between symbiotic
marine bacteria and their hosts is another field in which
proteomics may represent a valuable tool. In this sense,
2-D PAGE has also been successfully applied to the
elucidation of proteome changes in the squid Eupryma
scolopes due to the presence of the symbiotic marine
luminous bacterium Vibrio fischeri. These authors described the usefulness of 2-D PAGE in following up
protein changes in the host soluble proteome in colonized organs of the squid specimens in comparison with
uncolonized specimens.
10.11 Pharmacophore Model Hypo1, Virtual Screening for Identification
of Novel Tubulin Inhibitors with Potent Anticancer Activity
Microtubules are cytoskeletal filaments consisting of
R/ˇ-tubulin heterodimers and are involved in a wide
range of cellular functions, including shape maintenance, vesicle transport, cell motility, and division. In
the mitotic phase of the cell cycle, microtubules are in
dynamic equilibrium with tubulin dimers by assembling
the tubulin into microtubules or, conversely, disassembling microtubules to tubulin. Disruption of the dynamic
equilibrium can induce cell cycle arrest and ultimately
lead to apoptosis. Therefore, compounds that could in-
302 Part B Tools and Methods in Marine Biotechnology
other things, the improvement of the growth rate or
the increase in thermotolerance. Growth enhancement
strategies have been successfully applied to several
fish species, such as carp, catfish, tilapia, and salmon,
through the incorporation of homologous or heterologous growth hormone (GH) genes or cDNAs. The
monitoring of gene expression in genetically modified
marine organisms is thus a crucial step in the optimization of the use of such organisms in aquaculture
and seafood technology. It should also be noted that
the cross-effect of transgene expression on the biosynthesis of host proteins in genetically improved fish
species is another relevant issue in which proteomics
will surely play a significant role soon. In both issues,
proteomics represents a valuable tool of remarkable applied interest.
10.10 Present Status and Future Trends of Proteomics
in Marine Biotechnology
Proteomic tools, mainly 2-D PAGE (polyacrylamide
gel electrophoresis) and mass spectrometry, represent
a robust analytical strategy for the phenotypical investigation of marine organisms of industrial interest. These
tools afford specific information about expression patterns, post-translational modifications, or protein interaction mechanisms. This information is extremely
useful for the optimization of specific biotechnological processes. It should be stressed that high-throughput
proteomics techniques have recently been coupled with
newer bioinformatic tools, thus affording more robust
and powerful strategies for both protein identification and characterization purposes. Recently, proteomic
tools were successfully applied for the isolation and
characterization of a calponin-like protein from the
mussel Mytilus galloprovincialis 68 kDa polypeptide,
a polypeptide involved in muscle contraction processes.
The protein, isolated from the retractor muscle of this
mollusc, was successfully separated from actin by 2-D
PAGE and was identified by N-terminal amino acid
sequence analysis. Likewise, the investigation of mussel adhesive proteins which have been reported to
harbor an extraordinary biotechnological potential as
a biodegradable adhesive material is another promising field in which proteomics may play a significant
role. It is noteworthy that the protein can be readily obtained from mussels, a marine source characterized by
their wide availability and low price. Thus, proteomic
tools should play a significant role when undertaking
optimization strategies aimed at achieving the highest yields of such proteins, mainly by elucidating the
biochemical modifications to be considered in the process design step. The investigation of marine bacteria
by proteomic techniques is also still in its early days.
Although the protein profiles of the thermophilic microorganism Pyrococcus abyssi were investigated when
the organism was subjected to different levels of pressure and temperature, other authors applied 2-D PAGE
to investigate the level of protein biosynthesis and expression in Sphingomonas sp. cultivated under different
conditions. Earlier reports considered high-resolution
2-D PAGE to elucidate the mechanisms of UV protection in cyanobacteria, concluding that this technique
had greater power of resolution than conventional SDSPAGE (SDS: sodium dodecyl sulfate) to detect proteome changes. The relationship between symbiotic
marine bacteria and their hosts is another field in which
proteomics may represent a valuable tool. In this sense,
2-D PAGE has also been successfully applied to the
elucidation of proteome changes in the squid Eupryma
scolopes due to the presence of the symbiotic marine
luminous bacterium Vibrio fischeri. These authors described the usefulness of 2-D PAGE in following up
protein changes in the host soluble proteome in colonized organs of the squid specimens in comparison with
uncolonized specimens.
10.11 Pharmacophore Model Hypo1, Virtual Screening for Identification
of Novel Tubulin Inhibitors with Potent Anticancer Activity
Microtubules are cytoskeletal filaments consisting of
R/ˇ-tubulin heterodimers and are involved in a wide
range of cellular functions, including shape maintenance, vesicle transport, cell motility, and division. In
the mitotic phase of the cell cycle, microtubules are in
dynamic equilibrium with tubulin dimers by assembling
the tubulin into microtubules or, conversely, disassembling microtubules to tubulin. Disruption of the dynamic
equilibrium can induce cell cycle arrest and ultimately
lead to apoptosis. Therefore, compounds that could in-